Method of inhibiting matrix metalloproteinases

ABSTRACT

The present invention relates to a method of inhibiting matrix metalloproteinases using compounds that are dibenzofuran sulfonamide derivatives having the Formula I 
                 
 
     More particularly, the present invention relates to a method of treating diseases in which matrix metalloproteinases are involved such as multiple sclerosis, atherosclerotic plaque rupture, restenosis, aortic aneurism, heart failure, periodontal disease, corneal ulceration, burns, decubital ulcers, chronic ulcers or wounds, cancer metastasis, tumor angiogenesis, arthritis, or other autoimmune or inflammatory diseases dependent upon tissue invasion by leukocytes.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is a divisional application under 35 U.S.C. §121of U.S. patent application Ser. No. 10/162,518, filed Jun. 4, 2002, nowU.S. Pat. No. 6,620,835 which is a divisional application under 35U.S.C. §121 of U.S. patent application Ser. No. 09/254,384, filed Mar.2, 1999, now U.S. Pat. No. 6,624,177, which is a §371 of PCT/US97/14859,filed Aug. 22, 1997, which claims benefit of priority from U.S.Provisional Patent Application No. 60/025,062, filed Sep. 4, 1996, andNo. 60/055,713, filed Aug. 7, 1997.

FIELD OF THE INVENTION

The present invention relates to a method of inhibiting matrixmetalloproteinases using compounds that are dibenzofuran sulfonamidederivatives. More particularly, the present invention relates to amethod of treating diseases in which matrix metalloproteinases areinvolved such as multiple sclerosis, atherosclerotic plaque rupture,restenosis, aortic aneurism, heart failure, periodontal disease, cornealulceration, burns, decubital ulcers, chronic ulcers or wounds, cancermetastasis, tumor angiogenesis, arthritis, or other autoimmune orinflammatory diseases dependent upon tissue invasion by leukocytes.

BACKGROUND OF THE INVENTION

The compounds of the present invention are inhibitors of matrixmetalloproteinases, e.g., stromelysin-1 and gelatinase A (72 kDagelatinase).

Stromelysin-1 and gelatinase A are members of the matrixmetalloproteinases (MMP). Other members include fibroblast collagenase,neutrophil collagenase, gelatinase B (92 kDa gelatinase), stromelysin-2,stromelysin-3, matrilysin, collagenase 3, and the newly discoveredmembrane-associated matrix metalloproteinases (Sato H., Takino T., OkadaY., Cao J., Shinagawa A., Yamamoto E., and Seiki M., Nature,1994;370:61-65).

Stromelysin-1 is also known as MMP03 and gelatinase A is known as MMP02.In addition, several other matrix metalloproteinases are known:

-   -   MMP01—Fibroblast collagenase;    -   MMP07—Matrilysin;    -   MMP09 Gelatinase B; and    -   MMP13—Collagenase-3.

The catalytic zinc in matrix metalloproteinases is typically the focalpoint for inhibitor design. The modification of substrates byintroducing chelating groups has generated potent inhibitors such aspeptidehydroxamates and thiol-containing peptides. Peptide hydroxamatesand the natural endogenous inhibitors of MMPs (TIMPs) have been usedsuccessfully to treat animal models of cancer and inflammation.

The ability of the matrix metalloproteinases to degrade variouscomponents of connective tissue makes them potential targets forcontrolling pathological processes. For example, the rupture ofatherosclerotic plaques is the most common event initiating coronarythrombosis. Destabilization and degradation of the extracellular matrixsurrounding these plaques by MMPs has been proposed as a cause of plaquefissuring. The shoulders and regions of foam cell accumulation in humanatherosclerotic plaques show locally increased expression of gelatinaseB, stromelysin-1, and interstitial collagenase. In situ zymography ofthis tissue revealed increased gelatinolytic and caseinolytic activity(Galla Z. S., Sukhova G. K., Lark M. W., and Libby P., “Increasedexpression of matrix metalloproteinases and matrix degrading activity invulnerable regions of human atherosclerotic plaques”, J. Clin. Invest.,1994;94:2494-2503). In addition, high levels of stromelysin RNA messagehave been found to be localized to individual cells in atheroscleroticplaques removed from heart transplant patients at the time of surgery(Henney A. M., Wakeley P. R., Davies M. J., Foster K., Hembry R., MurphyG., and Humphries S., “Localization of stromelysin gene expression inatherosclerotic plaques by in situ hybridization,” Proc. Nat'l. Acad.Sci., 1991;88:8154-8158).

Inhibitors of matrix metalloproteinases will have utility in treatingdegenerative aortic disease associated with thinning of the medialaortic wall. Increased levels of the proteolytic activities of MMPs havebeen identified in patients with aortic aneurisms and aortic stenosis(Vine N. and Powell J. T., “Metalloproteinases in degenerative aorticdiseases,” Clin. Sci., 1991;81:233-239).

Heart failure arises from a variety of diverse etiologies, but a commoncharacteristic is cardiac dilation which has been identified as anindependent risk factor for mortality (Lee T. H., Hamilton M. A.,Stevenson L. W., Moriguchi J. D., Fonarow G. C., Child J. S., Laks H.,and Walden J. A., “Impact of left ventricular size on the survival inadvanced heart failure,” Am. J. Cardiol., 1993;72:672-676). Thisremodeling of the failing heart appears to involve the breakdown ofextracellular matrix. Matrix metalloproteinases are increased inpatients with both idiopathic and ischemic heart failure (Reddy H. K.,Tyagi S. C., Tjaha I. E., Voelker D. J., Campbell S. E., and Weber K.T., “Activated myocardial collagenase in idiopathic dilatedcardiomyopathy,” Clin. Res., 1993;41:660A; Tyagi S. C., Reddy H. K.,Voelker D., Tjara I. E., and Weber K. T., “Myocardial collagenase infailing human heart,” Clin. Res., 1993;41:681A). Animal models of heartfailure have shown that the induction of gelatinase is important incardiac dilation (Armstrong P. W., Moe G. W., Howard R. J., Grima E. A.,and Cruz T. F., “Structural remodeling in heart failure: gelatinaseinduction,” Can. J. Cardiol., 1994;10:214-220), and cardiac dilationprecedes profound deficits in cardiac function (Sabbah H. N., Kono T.,Stein P. D., Mancini G. B., and Goldstein S., “Left ventricular shapechanges during the course of evolving heart failure,” Am. J. Physiol.,1992;263:H266-H270). Neointimal proliferation, leading to restenosis,frequently develops after coronary angioplasty. The migration ofvascular smooth muscle cells (VSMCs) from the tunica media to theneointima is a key event in the development and progression of manyvascular diseases and a highly predictable consequence of mechanicalinjury to the blood vessel (Bendeck M. P., Zempo N., Clowes A. W.,Galardy R. E., and Reidy M., “Smooth muscle cell migration and matrixmetalloproteinase expression after arterial injury in the rat,”Circulation Research, 1994;75:539-545). Northern blotting andzymographic analyses indicated that gelatinase A was the principal MMPexpressed and excreted by these cells. Further, antisera capable ofselectively neutralizing gelatinase A activity also inhibited VSMCmigration across basement membrane barrier. After injury to the vessel,gelatinase A activity increased more than 20-fold as VSCMs underwent thetransition from a quiescent state to a proliferating, motile phenotype(Pauly R. R., Passaniti A., Bilato C., Monticone R., Cheng L.,Papadopoulos N., Gluzband Y. A., Smith L., Weinstein C., Lakatta E., andCrow M. T., “Migration of cultured vascular smooth muscle cells througha basement membrane barrier requires type IV collagenase activity and isinhibited by cellular differentiation,” Circulation Research,1994;75:41-54).

Collagenase and stromelysin activities have been demonstrated infibroblasts isolated from inflamed gingiva (Uitto V. J., Applegren R.,and Robinson P. J., “Collagenase and neutral metalloproteinase activityin extracts from inflamed human gingiva,” J. Periodontal Res.,1981;16:417-424), and enzyme levels have been correlated to the severityof gum disease (Overall C. M., Wiebkin O. W., and Thonard J. C.,“Demonstrations of tissue collagenase activity in vivo and itsrelationship to inflammation severity in human gingiva,” J. PeriodontalRes., 1987;22:81-88). Proteolytic degradation of extracellular matrixhas been observed in corneal ulceration following alkali burns (Brown S.I., Weller C. A., and Wasserman H. E., “Collagenolytic activity ofalkali burned corneas,” Arch. Opthalmol., 1969;81:370-373).Thiol-containing peptides inhibit the collagenase isolated fromalkali-burned rabbit corneas (Bums F. R., Stack M. S., Gray R. D., andPaterson C. A., Invest. Opththamol., 1989;30: 1569-1575).

Stromelysin is produced by basal keratinocytes in a variety of chroniculcers (Saarialho-Kere U. K., Ulpu K., Pentland A. P., Birkedal-HansenH., Parks W. C., Welgus H. G., “Distinct populations of basalkeratinocytes express stromelysin-1 and stromelysin-2 in chronicwounds,” J. Clin. Invest., 1994;94:79-88).

Stromelysin-1 mRNA and protein were detected in basal keratinocytesadjacent to but distal from the wound edge in what probably representsthe sites of proliferating epidermis. Stromelysin-1 may thus prevent theepidermis from healing. Davies, et al., (Cancer Res., 1993;53:2087-2091)reported that a peptide hydroxamate, BB-94, decreased the tumor burdenand prolonged the survival of mice bearing human ovarian carcinomaxenografts. A peptide of the conserved MMP propeptide sequence was aweak inhibitor of gelatinase A and inhibited human tumor cell invasionthrough a layer of reconstituted basement membrane (Melchiori A., AlbiliA., Ray J. M., and Stetler-Stevenson W. G., Cancer Res.,1992;52:2353-2356), and the natural tissue inhibitor ofmetalloproteinase-2 (TIMP-2) also showed blockage of tumor cell invasionin in vitro models (DeClerck Y. A., Perez N., Shimada H., Boone T. C.,Langley K. E., and Taylor S. M., Cancer Res., 1992;52:701-708). Studiesof human cancers have shown that gelatinase A is activated on theinvasive tumor cell surface (Strongin A. Y., Marmer B. L., Grant G. A.,and Goldberg G. I., J. Biol Chem., 1993;268:14033-14039) and is retainedthere through interaction with a receptor-like molecule (Monsky W. L.,Kelly T., Lin C.-Y., Yeh Y., Stetler-Stevenson W. G., Mueller S. C., andChen W.-T., Cancer Res., 1993;53:3159-3164). Inhibitors of MMPs haveshown activity in models of tumor angiogenesis (Taraboletti G., GarofaloA., Belotti D., Drudis T., Borsotti P., Scanziani E., Brown P. D., andGiavazzi R., Journal of the National Cancer Institute, 1995;87:293; andBenelli R., Adatia R., Ensoli B., Stetler-Stevenson W. G., Santi L., andAlbini A., Oncology Research, 1994;6:251-257).

Several investigators have demonstrated consistent elevation ofstromelysin and collagenase in synovial fluids from rheumatoid andosteoarthritis patients as compared to controls (Walakovits L. A., MooreV. L., Bhardwaj N., Gallick G. S., and Lark M. W., “Detection ofstromelysin and collagenase in synovial fluid from patients withrheumatoid arthritis and post-traumatic knee injury,” Arthritis Rheum.,1992;35:35-42; Zafarullah M., Pelletier J. P., Cloutier J. M., andMarcel-Pelletier J., “Elevated metalloproteinases and tissue inhibitorof metalloproteinase mRNA in human osteoarthritic synovia,” J.Rheumatol., 1993;20:693-697). TIMP-1 and TIMP-2 prevented the formationof collagen fragments, but not proteoglycan fragments, from thedegradation of both the bovine nasal and pig articular cartilage modelsfor arthritis, while a synthetic peptide hydroxamate could prevent theformation of both fragments (Andrews H. J., Plumpton T. A., Harper G.P., and Cawston T. E., Agents Actions, 1992;37:147-154; Ellis A. J.,Curry V. A., Powell E. K., and Cawston T. E., Biochem. Biophys. Res.Commun., 1994;201:94-101).

Gijbels, et al., (J. Clin. Invest., 1994;94:2177-2182) recentlydescribed a peptide hydroxamate, GM6001, that suppressed the developmentor reversed the clinical expression of experimental allergicencephalomyelitis (EAE) in a dose dependent manner, suggesting the useof MMP inhibitors in the treatment of autoimmune inflammatory disorderssuch as multiple sclerosis. A recent study by Madri has elucidated therole of gelatinase A in the extravasation of T-cells from the bloodstream during inflammation (Ramanic A. M. and Madri J. A., “TheInduction of 72-kD Gelatinase in T Cells upon Adhesion to EndothelialCells is VCAM-1 Dependent,” J. Cell Biology, 1994;125:1165-1178). Thistransmigration past the endothelial cell layer is coordinated with theinduction of gelatinase A and is mediated by binding to the vascularcell adhesion molecule-1 (VCAM-1). Once the barrier is compromised,edema and inflammation are produced in the CNS. Leukocytic migrationacross the blood-brain barrier is known to be associated with theinflammatory response in EAE. Inhibition of the metalloproteinasegelatinase A would block the degradation of extracellular matrix byactivated T-cells that is necessary for CNS penetration.

These studies provided the basis for the belief that an inhibitor ofstromelysin-1 and/or gelatinase A will treat diseases involvingdisruption of extracellular matrix resulting in inflammation due tolymphocytic infiltration, inappropriate migration of metastatic oractivated cells, or loss of structural integrity necessary for organfunction.

We have identified a series of tricyclic aromatic sulfonamide compoundsthat are inhibitors of matrix metalloproteinases, particularlystromelysin-1 and gelatinase A, and thus useful as agents for thetreatment of multiple sclerosis, atherosclerotic plaque rupture,restenosis, aortic aneurism, heart failure, periodontal disease, cornealulceration, burns, decubital ulcers, chronic ulcers or wounds, cancermetastasis, tumor angiogenesis, arthritis, or other autoimmune orinflammatory diseases dependent upon tissue invasion by leukocytes.

SUMMARY OF THE INVENTION

The present invention provides a method of inhibiting a matrixmetalloproteinase in a patient in need of matrix metalloproteinaseinhibition comprising administering to the patient a therapeuticallyeffective amount of a compound of Formula I

wherein M is a natural (L) alpha amino acid derivative having thestructure

-   X is O, S, S(O)_(n), CH₂, CO, or NR^(Q);-   R^(Q) is hydrogen, C₁-C₆ alkyl, or —C₁-C₆ alkyl-phenyl;-   R is a side chain of a natural alpha amino acid;-   R¹ is C₁-C₅ alkoxy, hydroxy, or —NHOR⁵;-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl -NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides and prodrugs thereof.

In one embodiment of the invention of Formula I, X is O.

In another embodiment of the invention of Formula I, X is S.

In another embodiment of the invention of Formula I, X is CH₂.

In another embodiment of the invention of Formula I, X is NR^(Q).

In a preferred embodiment of the invention of Formula I, X is O and R²and R⁴ are hydrogen.

In another embodiment of the invention of Formula I, X is CO.

In another embodiment of the invention of Formula I, X is S(O)_(n).

In another preferred embodiment of the invention of Formula I, R¹ ishydroxy, C₁-C₅ alkoxy, —NHOH, or —NHO benzyl.

In still another preferred embodiment, R is the side chain of thenatural alpha amino acid glycine, alanine, valine, leucine, isoleucine,cysteine, aspartic acid, or phenylalanine.

In another embodiment, the present invention provides a method ofinhibiting a matrix metalloproteinase in a patient in need of matrixmetalloproteinase inhibition, the method comprising administering to thepatient a therapeutically effective amount of a compound of Formula II

wherein Z is a natural (L) amino acid derivative having the structure

-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR^(c);-   R^(b) is a side chain of a natural alpha amino acid; and-   R^(c) is hydrogen, C₁-C₅ alkyl, or —CH₂ phenyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

In a preferred embodiment of the method comprising Formula II, the group

is located at the 2-position of the phenyl ring.

In another preferred embodiment of the method comprising Formula II, thegroup

is located at the 3-position of the phenyl ring.

Also provided is a method of inhibiting a matrix metalloproteinase in apatient in need of matrix metalloproteinase inhibition, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of Formula III

wherein Z is a natural (L) amino acid derivative having the structure

-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR^(c);-   R^(b) is a side chain of a natural alpha amino acid; and-   R^(c) is hydrogen, C₁-C₅ alkyl, or —CH₂ phenyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

Also provided is a method of inhibiting a matrix metalloproteinase in apatient in need of matrix metalloproteinase inhibition, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of Formula IV.

wherein Z is a natural (L) amino acid derivative having the structure

-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR^(c);-   R^(b) is a side chain of a natural alpha amino acid; and-   R^(c) is hydrogen, C₁-C₅ alkyl, or —CH₂ phenyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

Also provided is a method of inhibiting a matrix metalloproteinase in apatient in need of matrix metalloproteinase inhibition, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of Formula V

wherein Z is a natural (L) amino acid derivative having the structure

-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR^(c);-   R^(b) is a side chain of a natural alpha amino acid; and-   R^(c) is hydrogen, C₁-C₅ alkyl, or —CH₂ phenyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

Also provided is a method of inhibiting a matrix metalloproteinase in apatient in need of matrix metalloproteinase inhibition, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of Formula VI

wherein Z is a natural (L) amino acid derivative having the structure

-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR^(c);-   n is 0 to 2;-   R^(b) is a side chain of a natural alpha amino acid; and-   R^(c) is hydrogen, C₁-C₅ alkyl, or —CH₂ phenyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

Also provided is a method of inhibiting a matrix metalloproteinase in apatient in need of matrix metalloproteinase inhibition, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of Formula VII

wherein Z is a natural (L) amino acid derivative having the structure

-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR^(c);-   R^(Q) is hydrogen, C₁-C₆ alkyl, or C₁-C₆ alkyl-phenyl;-   R^(b) is a side chain of a natural alpha amino acid; and-   R^(c) is hydrogen, C₁-C₅ alkyl, or —CH₂ phenyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

In a most preferred embodiment, the compound of Formula I-VIII is:

-   -   (L)-2-(dibenzofuran-2-sulfonylamino)-4-methyl-pentanoic acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-phenyl-propionic acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-propionic acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-methyl-butyric acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-acetic acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-succinic acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-tritylsulfanyl-propionic        acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-mercapto-propionic acid;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acid        hydroxyamide;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-acetic acid tert-butyl        ester;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-propionic acid tert-butyl        ester;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-propionic acid tert-butyl        ester;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-4-methyl-pentanoic acid        tert-butyl ester;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acid        tert-butyl ester;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acid        benzyloxy-amide;    -   (L)-2-(dibenzofuran-2-sulfonylamino)-3-phenyl-propionic acid        tert-butyl ester;    -   (L)-2-(dibenzofuran-3-sulfonylamino)-3-methyl-butyric acid;    -   3-Methyl-2-(9-methyl-9H-carbazole-3-sulfonylamino)-butyric acid;    -   2-(9-Benzyl-9H-carbazole-3-sulfonylamino)-3-methyl-butyric acid;    -   (L)-2-(9H-Fluorene-2-sulfonylamino)-3-methyl-butyric acid;    -   (L)-2-(5,5-Dioxo-5H-5λ⁶-dibenzothiophene-3-sulfonylamino)-3-methyl-butyric        acid;    -   (L)-2-(Dibenzothiophene-2-sulfonylamino)-3-methyl-butyric acid;    -   (L)-2-(7-Bromo-dibenzofuran-2-sulfonylamino)-3-methyl-butyric        acid;    -   (L)-3-Methyl-2-(7-phenyl dibenzofuran-2-sulfonylamino)-butyric        acid; and    -   2-(9H-Carbazole-3-sulfonylamino)-3-methyl-butyric acid.

Also provided by the present invention is a method of treating multiplesclerosis, the method comprising administering to a patient havingmultiple sclerosis a therapeutically effective amount of a compound ofFormula I-VIII.

Also provided by the present invention is a method of treatingatherosclerotic plaque rupture, the method comprising administering to apatient having an atherosclerotic plaque at risk for rupture atherapeutically effective amount of a compound of Formula I-VIII.

Also provided by the present invention is a method of treating orpreventing restenosis, the method comprising administering to a patienthaving restenosis or at risk of having restenosis a therapeuticallyeffective amount of a compound of Formula I-VIII.

Also provided by the present invention is a method of treating aorticaneurism, the method comprising administering to a patient having aorticaneurism a therapeutically effective amount of a compound of FormulaI-VIII.

Also provided by the present invention is a method of treating heartfailure, the method comprising administering to a patient having heartfailure a therapeutically effective amount of a compound of FormulaI-VIII.

Also provided by the present invention is a method of treatingperiodontal disease, the method comprising administering to a patienthaving periodontal disease a therapeutically effective amount of acompound of Formula I-VIII.

Also provided by the present invention is a method of treating cornealulceration, the method comprising administering to a patient havingcorneal ulceration a therapeutically effective amount of a compound ofFormula I-VIII.

Also provided by the present invention is a method of treating burns,the method comprising administering to a patient having burns atherapeutically effective amount of a compound of Formula I-VIII.

Also provided by the present invention is a method of treating decubitalulcers, the method comprising administering to a patient havingdecubital ulcers a therapeutically effective amount of a compound ofFormula I-VIII.

Also provided by the present invention is a method of treating chroniculcers or wounds, the method comprising administering to a patienthaving chronic ulcers or wounds a therapeutically effective amount of acompound of Formula I-VIII.

Also provided by the present invention is a method of treating cancermetastasis, the method comprising administering to a patient havingcancer metastasis a therapeutically effective amount of a compound ofFormula I-VIII.

Also provided by the present invention is a method of treating tumorangiogenesis, the method comprising administering to a patient havingtumor angiogenesis a therapeutically effective amount of a compound ofFormula I-VIII.

Also provided by the present invention is a method of treatingarthritis, the method comprising administering to a patient havingarthritis a therapeutically effective amount of a compound of FormulaI-VIII.

Also provided by the present invention is a method of treatingautoimmune or inflammatory diseases dependent upon tissue invasion byleukocytes, the method comprising administering to a patient havingautoimmune or inflammatory diseases dependent upon tissue invasion byleukocytes a therapeutically effective amount of a compound of FormulaI-VIII.

The present invention also provides compounds of Formula I

wherein M is a natural (L) alpha amino acid derivative having thestructure

-   X is S, S(O)_(n), CH₂, CO, or NR^(Q);-   R^(b) is a side chain of a natural alpha amino acid;-   R^(Q) is hydrogen, C₁-C₆ alkyl, or —C₁-C₆ alkyl-phenyl;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR⁵;-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

The present invention also provides compounds of Formula VIII

wherein M is a natural (L) alpha amino acid derivative having thestructure

-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   R^(b) is a side chain of a natural alpha amino acid;-   R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR⁵;-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a method of inhibiting a matrixmetalloproteinase in a patient in need of matrix metalloproteinaseinhibition comprising administering to the patient a therapeuticallyeffective amount of a compound of Formula I

wherein M is a natural (L) alpha amino acid derivative having thestructure

-   X is O, S, S(O)_(n), CH₂, CO, or NR^(Q);-   R is a side chain of a natural alpha amino acid;-   R¹ is C₁-C₅ alkoxy, hydroxy, or —NHOR⁵;-   R² and R⁴ are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂,    halogen, —OR⁵, —CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶,    —(CH₂)_(n)NR⁵R⁶, —CF₃, or —NHCOR⁵;-   each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl; and-   n is 0 to 2, and the pharmaceutically acceptable salts, esters,    amides, and prodrugs thereof.

The term “alkyl” means a straight or branched chain hydrocarbon.Representative examples of alkyl groups are methyl, ethyl, propyl,isopropyl, isobutyl, butyl, tert-butyl, sec-butyl, pentyl, and hexyl.

The term “alkoxy” means an alkyl group attached to an oxygen atom.Representative examples of alkoxy groups include methoxy, ethoxy,tert-butoxy, propoxy, and isobutoxy.

The term “halogen” includes chlorine, fluorine, bromine, and iodine.

The term “phenyl” also includes substituted phenyl wherein one or morehydrogen on the phenyl ring is replaced with an organic radical.Examples of suitable substituents include, but are not limited to,halogen, C₁-C₆ alkoxy, —CF₃, —NO₂, —NH₂, —NH(C₁-C₆ alkyl), or —N(C₁-C₆alkyl)₂.

The symbol “-” means a bond.

The term “side chain of a natural alpha amino acid” means the group Q ina natural amino acid of formula H₂N—CH(Q)—COOH. Examples of side chainsof natural alpha amino acids include those of alanine, arginine,asparagine, aspartic acid, cysteine, glutamic acid, glycine, histidine,isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine,threonine, tryptophan, tyrosine and valine.

A natural alpha amino acid is an amino acid found in a living organism.Examples of such amino acids include glycine, alanine, valine, leucine,isoleucine, phenylalanine, proline, serine, threonine, tyrosine,asparagine, glutamine, lysine, arginine, tryptophan, histidine,cysteine, methionine, aspartic acid, and glutamic acid.

The functional groups in the amino acid side chains can be protected.For example, carboxyl groups can be esterified, amino groups can beconverted to amides or carbamates, hydroxyl groups can be converted toethers or esters, and thiol groups can be converted to thioethers orthioesters.

The compounds of Formula I-VIII can be administered to a patient eitheralone or as part of a pharmaceutically acceptable composition. Thecompositions can be administered to patients such as humans and animalseither orally, rectally, parenterally (intravenously, intramuscularly orsubcutaneously), intracisternally, intravaginally, intraperitoneally,intravesically, locally (powders, ointments or drops), or as a buccal ornasal spray.

Compositions suitable for parenteral injection may comprisephysiologically acceptable sterile aqueous or nonaqueous solutions,dispersions, suspensions or emulsions, and sterile powders forreconstitution into sterile injectable solutions or dispersions.Examples of suitable aqueous and nonaqueous carriers, diluents,solvents, or vehicles include water, ethanol, polyols (propyleneglycol,polyethyleneglycol, glycerol, and the like), suitable mixtures thereof,vegetable oils (such as olive oil), and injectable organic esters suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preserving,wetting, emulsifying, and dispensing agents. Prevention of the action ofmicroorganisms can be ensured by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, sorbic acid, andthe like. It may also be desirable to include isotonic agents, forexample sugars, sodium chloride, and the like. Prolonged absorption ofthe injectable pharmaceutical form can be brought about by the use ofagents delaying absorption, for example, aluminum monostearate andgelatin.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is admixed with at least one inert customary excipient (orcarrier) such as sodium citrate or dicalcium phosphate or (a) fillers orextenders, as for example, starches, lactose, sucrose, glucose,mannitol, and silicic acid, (b) binders, as for example,carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone,sucrose, and acacia, (c) humectants, as for example, glycerol, (d)disintegrating agents, as for example, agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, certain complex silicates, andsodium carbonate, (e) solution retarders, as for example paraffin, (f)absorption accelerators, as for example, quaternary ammonium compounds,(g) wetting agents, as for example, cetyl alcohol and glycerolmonostearate, (h) adsorbents, as for example, kaolin and bentonite, and(i) lubricants, as for example, talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate or mixturesthereof. In the case of capsules, tablets, and pills, the dosage formsmay also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethyleneglycols, andthe like.

Solid dosage forms such as tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells, such as entericcoatings and others well-known in the art. They may contain opacifyingagents, and can also be of such composition that they release the activecompound or compounds in a certain part of the intestinal tract in adelayed manner. Examples of embedding compositions which can be used arepolymeric substances and waxes. The active compounds can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirs. Inaddition to the active compounds, the liquid dosage forms may containinert diluents commonly used in the art, such as water or othersolvents, solubilizing agents and emulsifiers, as for example, ethylalcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol,dimethylformamide, oils, in particular, cottonseed oil, groundnut oil,corn germ oil, olive oil, castor oil and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethyleneglycols, and fatty acid estersof sorbitan or mixtures of these substances, and the like.

Besides such inert diluents, the composition can also include adjuvants,such as wetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspendingagents, as for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, or mixtures of thesesubstances, and the like.

Compositions for rectal administrations are preferably suppositorieswhich can be prepared by mixing the compounds of the present inventionwith suitable non-irritating excipients or carriers such as cocoabutter, polyethyleneglycol, or a suppository wax, which are solid atordinary temperatures but liquid at body temperature and therefore, meltin the rectum or vaginal cavity and release the active component.

Dosage forms for topical administration of a compound of this inventioninclude ointments, powders, sprays, and inhalants. The active componentis admixed under sterile conditions with a physiologically acceptablecarrier and any preservatives, buffers, or propellants as may berequired. Ophthalmic formulations, eye ointments, powders, and solutionsare also contemplated as being within the scope of this invention.

The compounds of the present invention can be administered to a patientat dosage levels in the range of about 0.1 to about 1,000 mg per day.For a normal human adult having a body weight of about 70 kg, a dosagein the range of about 0.01 to about 100 mg/kg of body weight per day ispreferable. The specific dosage used, however, can vary. For example,the dosage can depend on a numbers of factors including the requirementsof the patient, the severity of the condition being treated, and thepharmacological activity of the compound being used. The determinationof optimum dosages for a particular patient is well-known to thoseskilled in the art. The term “patient” includes humans and animal.

The term “pharmaceutically acceptable salts, esters, amides, andprodrugs” as used herein refers to those carboxylate salts, amino acidaddition salts, esters, amides, and prodrugs of the compounds of thepresent invention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of patients without unduetoxicity, irritation, allergic response, and the like, commensurate witha reasonable benefit/risk ratio, and effective for their intended use,as well as the zwitterionic forms, where possible, of the compounds ofthe invention. The term “salts” refers to the relatively nontoxic,inorganic and organic acid addition salts of compounds of the presentinvention. These salts can be prepared in situ during the finalisolation and purification of the compounds or by separately reactingthe purified compound in its free base form with a suitable organic orinorganic acid and isolating the salt thus formed. Representative saltsinclude the hydrobromide, hydrochloride, sulfate, bisulfate, nitrate,acetate, oxalate, valerate, oleate, palmitate, stearate, laureate,borate, benzoate, lactate, phosphate, tosylate, citrate, maleate,fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate,lactobionate and laurylsulphonate salts, and the like. These may includecations based on the alkali and alkaline earth metals, such as sodium,lithium, potassium, calcium, magnesium, and the like, as well asnontoxic ammonium, quaternary ammonium, and amine cations including, butnot limited to ammonium, tetramethylammonium, tetraethylammonium,methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine,and the like. (See, for example, S. M. Berge, et al., “PharmaceuticalSalts,” J. Pharm. Sci., 1977:66(1-19) which is incorporated herein byreference.)

Examples of pharmaceutically acceptable, nontoxic esters of thecompounds of this invention include C₁ to C₆ alkyl esters wherein thealkyl group is a straight or branched chain. Acceptable esters alsoinclude C₅ to C₇ cycloalkyl esters as well as arylalkyl esters such as,but not limited to benzyl. C₁ to C₄ alkyl esters are preferred. Estersof the compounds of the present invention may be prepared according toconventional methods.

Examples of pharmaceutically acceptable, nontoxic amides of thecompounds of this invention include amides derived from ammonia, primaryC₁ to C₆ alkyl amines, and secondary C₁ to C₆ dialkyl amines wherein thealkyl groups are straight or branched chain. In the case of secondaryamines, the amine may also be in the form of a 5- or 6-memberedheterocycle containing one nitrogen atom. Amides derived from ammonia,C₁ to C₃ alkyl primary amines and C₁ to C₂ dialkyl secondary amines arepreferred. Amides of the compounds of the invention may be preparedaccording to conventional methods.

The term “prodrug” refers to compounds that are rapidly transformed invivo to yield the parent compound of the above formula, for example, byhydrolysis in blood. A thorough discussion is provided in T. Higuchi andV. Stella, “Pro-drugs as Novel Delivery Systems,” Vol 14 of the A.C.S.Symposium Series, and in Bioreversible Carriers in Drug Design, ed.Edward B. Roche, American Pharmaceutical Association and Pergamon Press,1987, both of which are incorporated herein by reference.

In addition, the compounds of the present invention can exist inunsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms for the purposesof the present invention.

The compounds of the present invention are administered to a patient inneed of matrix metalloproteinase inhibition. In general, patients inneed of matrix metalloproteinase inhibition are those patients having adisease or condition in which a matrix metalloproteinase plays a role.Examples of such diseases include, but are not limited to, multiplesclerosis, atherosclerotic plaque rupture, restenosis, aortic aneurism,heart failure, periodontal disease, corneal ulceration, burns, decubitalulcers, chronic ulcers or wounds, cancer metastasis, tumor angiogenesis,arthritis, or other autoimmune or inflammatory diseases dependent upontissue invasion by leukocytes.

In a preferred embodiment, the matrix metalloproteinase is stromelysin-1or gelatinase-A.

A “therapeutically effective amount” is an amount of a compound ofFormula I-VIII that when administered to a patient having a disease thatcan be treated with a compound of Formula I-VIII ameliorates a symptomof the disease. A therapeutically effective amount of a compound ofFormula I-VIII is readily determined by one skilled in the art byadministering a compound of Formula I-VIII to a patient and observingthe results.

The following examples illustrate particular embodiments of theinvention and are not intended to limit the scope of the specification,including the Claims, in any manner.

EXAMPLES

General Dibenzofuran Sulfonamide Synthesis

The compounds of the present invention can be synthesized using a numberof different synthetic routes. Referring to the General SyntheticScheme, the common starting materials are the sulfonyl chlorides (1).These are easily synthesized by one skilled in the art by sulfonation ofthe parent heterocycle. Some representative procedures are as follows.For dibenzofuran (1, X═O) and dibenzothiophene (1, X═S), the parentheterocycle is sulfonated at the 2-position using one equivalent ofchlorosulfonic acid in chloroform at 0° C. according to the method ofBassin, et al., (Phosphorus, Sulfur and Silicon, 1992;72:157-170). Thesulfonic acid is then converted to the corresponding sulfonyl chloride(1, X═O,S) by treatment with phosphorus pentachloride at 170-180° C. Forcarbazole (1, X═NH), the parent heterocycle is sulfonated at the3-position using sulfuric acid at 100° C. followed by neutralizationwith barium carbonate to yield the barium salt of the correspondingsulfonic acid according to the method of Loza, et al., (Sb. Mater.Nauch.-Tekh. Konf. Ukrain. Zaoch. Poitekh. Inst. Vith, Kharkov,1966:202-205). The sulfonic acid is then converted to the correspondingsulfonyl chloride (1, X═NH) by treatment with phosphorus pentachlorideat 170-180° C. or reaction with either phosphoryl chloride, thionylchloride, or oxalyl chloride. For fluorene (1, X═CH₂), according to themethod of Chrzaszczewska et al., (Lodz. Tow. Nauk., Wydz. 3, Acta Chim.,1966;11: 143-155) the parent carbocycle is sulfonated at the 2-positionusing one equivalent of chlorosulfonic acid in chloroform at 0° C.followed by neutralization with potassium hydroxide to give thepotassium salt of the corresponding sulfonic acid. This fluorenederivative can then be oxidized using aqueous potassium permanganate at80° C. to the corresponding fluorenone derivative (1, X═CO). Thesulfonic acid salts are then converted to the corresponding sulfonylchloride (1, X═CH₂,CO) by treatment with phosphorus pentachloride andphosphoryl chloride in chloroform.

In Method A, the sulfonyl chloride (1) is condensed directly with anatural amino acid using a base such as triethylamine (TEA) in a mixtureof tetrahydrofuran (THF) and water (3:5) at 10° C. to yield the desiredcompound (2). The corresponding hydroxamic acid (5) can be convenientlyprepared by coupling the acid (2) with an O-protected (usually benzyl)hydroxylamine using dicyclohexylcarbodiimide (DCC) as the coupling agentin dichloromethane at temperatures ranging from −(10) to 0° C. Theprotecting group can be removed from compound (4) by catalytichydrogenolysis using hydrogen gas at 50 psi and Pd/BaSO₄ in aqueousmethanol to yield the hydroxamic acid derivative (5).

In Method B, the sulfonyl chloride (1) is condensed with a suitablyC-protected (usually tertiary butyl ester) amino acid using a base suchas N-methylmorpholine (NMM) in a solvent such as dichloromethane at 0°C. to yield compound (3). The protecting group can be removed from thecarboxylic acid by treatment with trifluoroacetic acid indichloromethane at 25-35° C. using anisole as a carbocation scavenger toyield (2).

Examples Prepared by Method A

Example 1 (L)-2-(Dibenzofuran-2-sulfonylamino)-4-methyl-pentanoic acid

Step (a) (L)-2-(Dibenzofuran-2-sulfonylamino)-4-methyl-pentanoic acid,tert.-butyl ester

To a dichloromethane solution (20 mL) of (L)-leucine, tert.-butyl ester(2.1 g, 0.0099 mol) and N-methylmorpholine (2.2 mL, 0.0199 mol) at 0° C.under an inert nitrogen atmosphere was added a dichloromethane solution(10 mL) of dibenzofuran-2-sulfonyl chloride (1.0 g, 0.00375 mol) withstirring. The resulting solution was stirred at 0° C. for 4 hours andthen partitioned with water (30 mL). The organic layer was separated andwashed with water (2×30 mL) and brine (2×30 mL). This was then driedover anhydrous magnesium sulfate, filtered, and the solvent removedunder reduced pressure. The residue was then flash chromatographed onsilica gel and the title product (1.0 g, 64%) was eluted with 20% ethylacetate/hexane; melting point=106-109° C.

Step (b) (L)-2-(Dibenzofuran-2-sulfonylamino)-4-methyl-pentanoic acid

Example 1

To a dichloromethane solution (5 mL) of the material obtained in step(a) (0.5 g, 0.00119 mol) and anisole (0.5 mL) at room temperature withstirring was added trifluoroacetic acid (5 mL). The resulting solutionwas stirred at room temperature for 24 hours and then concentrated invacuo. The residue was triturated with a mixture of ethyl acetate/hexaneto yield the title compound (0.14 g, 33%); melting point=75-80° C.

¹H NMR (CDCl₃: Λ 8.4 (s, 1H), 8.0 (d, 1H), 7.9 (d, 1H), 7.4-7.6 (m, 4H),5.0 (d, 1H), 3.9 (m, 1H), 1.8 (m, 1H), 1.4 (m, 2H), 0.9 (d, 3H), 0.8 (d,3H) ppm.

Following the general procedure of Example 1, the following compoundswere obtained:

Example 2

(L)-2-(Dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acid

¹H NMR (DMSO-D₆): Λ 8.6 (s, 1H), 8.3 (d, 1H), 8.1 (d, 1H), 7.8-7.9 (m,3H), 7.6 (tr, 1H), 7.5 (tr, 1H), 3.7 (m, 1H), 3.4 (s, 1H), 1.7 (m, 1H),1.1-1.4 (m, 2H), 0.75-0.85 (m, 6H) ppm.

Example 3

(L)-2-(Dibenzofuran-2-sulfonylamino)-3-phenyl-propionic acid; MeltingPoint=196-198° C.

Examples Prepared by Method B

Example 4

(L)-2-(Dibenzofuran-2-sulfonylamino)-propionic acid

To a THF/water (5:3, 8 mL) solution of (L)-alanine (0.3 g, 0.0034 mol)and triethylamine (1 mL) at 10° C. was added dibenzofuran-2-sulfonylchloride (1.0 g, 0.00375 mol) in one portion with stirring. Theresulting solution was stirred at room temperature for 24 hours. Thesolution was then concentrated in vacuo and the residue redissolved inwater (10 mL). This solution was cooled in an ice bath and thenacidified with 1N HCl. A white solid was deposited which was thenfiltered and washed with water. This solid was recrystallized fromaqueous ethanol to give the title product (0.6 g, 50%); meltingpoint=158-163° C.

Following the general procedure of Example 4, the following compoundswere obtained:

Example 5

(L)-2-(Dibenzofuran-2-sulfonylamino)-3-methyl-butyric acid; meltingPoint=163-165° C.

Example 6

(Dibenzofuran-2-sulfonylamino)-acetic acid; Melting Point=208-210° C.

Example 7

(L)-2-(Dibenzofuran-2-sulfonylamino)-succinic acid; meltingpoint=165-168° C.

Example 8

(L)-2-(Dibenzofuran-2-sulfonylamino)-3-tritylsulfanyl-propionic acid

¹NMR (DMSO-D₆): Λ 8.5 (s, 1H), 8.2 (m, 2H), 7.1-7.9 (m, 19H), 3.6 (m,1H), 3.5 (m, 1H), 2.3 (d, 2H) ppm.

Example 9

(L)-2-(Dibenzofuran-2-sulfonylamino)-3-mercapto-propionic acid

To a dichloromethane solution (10 mL) of(L)-2-(Dibenzofuran-2-sulfonylamino)-3-tritylsulfanyl-propionic acid(Example 8, 1.0 g, 0.00168 mol) at room temperature was addedtrifluoroacetic acid (10 mL). A deep red/orange solution resulted. Tothis solution was added triethylsilane (0.33 mL, 0.00202 mol), the colorwas immediately discharged, and the resulting clear solution was stirredat room temperature for 3 hours. The solution was then concentrated invacuo and the residue redissolved in ether (10 mL) which was thenremoved in vacuo. This procedure was repeated three times. The residuewas recrystallized from ethyl acetate/hexane (1:1) to yield the titlecompound (0.23 g, 39%); melting point=164-166° C.

Example 10

(L)-2-(Dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acidhydroxyamide

Step (a) (L)-2-(Dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acidbenzyloxy-amide

To a THF solution (50 mL) of(L)-2-(Dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acid (Example 2,0.55 g, 0.0015 mol) and carbonyldiimidazole (0.26 g, 0.0016 mol) at roomtemperature under an inert nitrogen atmosphere was addedO-benzylhydroxylamine (0.23 g, 0.0018 mol) in one portion. This solutionwas then heated to reflux for 72 hours and then allowed to stir at roomtemperature for 24 hours. The mixture was then concentrated in vacuo andflash chromatographed on silica gel eluting with ethyl acetate/hexane(1:4) to yield the title compound (0.27 g, 38%); melting point=207-209°C.

Step (b) (L)-2-(Dibenzofuran-2-sulfonylamino)-3-methyl-pentanoic acidhydroxyamide

Example 10

A THF (2 mL)/methanol (10 mL) solution of the material obtained above instep (a) (0.037 g, 0.0000793 mol) was hydrogenolyzed using hydrogen gasat 50 psi with a Pd/BaSO₄ catalyst at room temperature for 1 hour. Thecatalyst was removed by filtration and the solution concentrated invacuo. The residue was triturated with ether to yield the title compound(0.022 g, 74%).

¹H NMR (CDCl₃): Λ 8.6-7.2 (m, 8H), 5.1 (m, 1H), 4.1 (m, 1H), 1.9-1.2 (m,3H), 0.9 (m, 3H), 0.85 (m, 3H) ppm.

Example 11

(L)-2-(dibenzofuran-3-sulfonylamino)-3-methyl-butyric acid

Step (a) (Dibenzofuran-3-sulfonyl chloride)

3-Aminodibenzofuran (10 g, 54.6 mol) was diazotized by dissolving in 180mL glacial acetic acid, 50 mL water, and 14 mL concentrated hydrochloricacid at 0° C. and adding 15 mL of a 5.5 M aqueous solution of sodiumnitrite. The resulting mixture was stirred for 1 hour before pouringinto a solution of copper(II)chloride (2.0 g, 14.9 mmol) in 240 mL of a1:1 mixture of benzene and glacial acetic acid saturated with sulfurdioxide. This mixture was allowed to warm to room temperature andstirred for 16 hours. The reaction was partitioned between water andchloroform. The chloroform layer was washed with water, dried overmagnesium sulfate, filtered, and concentrated to give the title compoundas a yellowish solid; melting point=142-144° C.

Step (b)

Using the procedure of Example 1, (L)-leucine, tert.-butyl ester isreplaced with (L)-valine, tert.-butyl ester and dibenzofuran-2-sulfonylchloride is replaced with dibenzofuran-3-sulfonyl chloride, the titlecompound is obtained; melting point=197-200° C.

Example 12

(L)-2-(9H-Fluorene-2-sulfonylamino)-3-methyl-butyric acid

When in the procedure of EXAMPLE 1, (L)-leucine, tert.-butyl ester isreplaced with (L)-valine, tert.-butyl ester and dibenzofuran-2-sulfonylchloride is replaced with 9H-fluorene-2-sulfonyl chloride, the titlecompound is obtained.

Example 13

(L)-2-(5,5-Dioxo-5H-5λ⁶-dibenzothiophene-3-sulfonylamino)-3-methyl-butyricacid

When in the procedure of EXAMPLE 1, (L)-leucine, tert.-butyl ester isreplaced with (L)-valine, tert.-butyl ester and dibenzofuran-2-sulfonylchloride is replaced with 5,5-dioxo-5H-5λ⁶-dibenzothiophene-3-sulfonylchloride, the title compound is obtained; melting point=85-90° C.

Example 14

(L)-2-(Dibenzothiophene-2-sulfonylamino)-3-methyl-butyric acid

When in the procedure of EXAMPLE 1, (L)-leucine, tert.-butyl ester isreplaced with (L)-valine, tert.-butyl ester and dibenzofuran-2-sulfonylchloride is replaced with dibenzothiophene-2-sulfonyl chloride, thetitle compound is obtained; melting point=150-155° C.

Example 15

(L)-2-(5,5-Dioxo-5H-5λ⁶-dibenzothiophene-2-sulfonylamino)-3-methyl-butyricacid

To a glacial acetic acid (30 mL) solution of the material obtained inExample 14 Step (a) (1.5 g, 0.0036 mol) was added 10 mL of 30% hydrogenperoxide. The resulting solution was heated to reflux for 2.5 hours,cooled to room temperature and stirred for 16 hours and then filtered togive the crude product as a white solid. The solid was washed with waterand boiling ether to yield the title compound; melting point=216-218° C.

Example 16

(L)-2-(7-Bromo-dibenzofuran-2-sulfonylamino)-3-methyl-butyric acid

Step (a) 3-Bromo-dibenzofuran

3-Amino-dibenzofuran (15 g, 81.9 mmoles) was added in portions to asuspension of cupric bromide (21.9 g, 98.2 mmoles) and tert.-butylnitrite (12.66 g, 122.8 mmoles) in 350 mL of acetonitrile. This mixturewas heated to reflux for 2 hours and then stirred for 16 hours at roomtemperature. The reaction was partitioned between 1 M HCl and diethylether. The diethyl ether layer was washed with brine, dried overmagnesium sulfate, filtered, and concentrated to give an oily solid.Chromatography gave the title compound as a yellowish solid.

Step (b) 7-Bromo-dibenzofuran-2-sulfonyl chloride

Chlorosulfonic acid (3.75 mL, 56 mmoles) was added dropwise to asolution of 3-bromo-dibenzofuran (9.21 g, 37.3 mmoles) in 150 mL ofchloroform at room temperature. The reaction was stirred for 5 hours,cooled to 0° C., filtered, and washed the solid with colddichloromethane. This solid (6.12 g, 18.7 mmoles) was mixed withphosphorous pentachloride (12.9 g, 61.7 mmoles) and the mixture washeated to 110° C. for 4 hours. The mixture was cooled to roomtemperature and quenched with ice water. Filtered the resultingsuspension to give the title compound as a white solid.

Step (c) (L)-2-(7-Bromo-dibenzofuran-2-sulfonylamino)-3-methyl-butyricacid

When in the procedure of EXAMPLE 1, (L)-leucine, tert.-butyl ester isreplaced with (L)-valine, tert.-butyl ester and dibenzofuran-2-sulfonylchloride is replaced with 7-bromo-dibenzofuran-2-sulfonyl chloride, thetitle compound is obtained; melting point=191-193° C.

Example 17

(L)-3-Methyl-2-(7-phenyl-dibenzofuran-2-sulfonylamino)-butyric acid

Step (a) (L)-2-(7-Bromo-dibenzofuran-2-sulfonylamino)-butyric acid,tert.-butyl ester

When in the procedure of EXAMPLE 1, Step (a), (L)-leucine, tert.-butylester is replaced with (L)-valine, tert.-butyl ester anddibenzofuran-2-sulfonyl chloride is replaced with7-bromo-dibenzofuran-2-sulfonyl chloride, the title compound isobtained.

Step (b) (L)-3-Methyl-2-(7-phenyl-dibenzofuran-2-sulfonylamino)-butyricacid, tert.-butyl ester

(L)-2-(7-Bromo-dibenzofuran-2-sulfonylamino)-3-methyl-butyric acid,tert.-butyl ester (1.0 g, 2.0 mmoles) and phenyl boronic acid (0.3 g,2.5 mmoles) were mixed with 10 mL toluene with 5 mL water the 0.5 gsodium carbonate. Tetrakis(triphenylphosphine) palladium (0) (0.15 g,0.1 mmoles) was added and the resulting mixture was heated to reflux for6 hours. Another 0.15 g of the palladium catalyst was added and refluxwas continued for 16 hours. The reaction was cooled to room temperatureand partitioned between 1 M HCl and ethyl acetate. The organic layer wasdried over magnesium sulfate and concentrated to give the title compoundas a white solid.

Step (c) (L)-3-Methyl-2-(7-phenyl-dibenzoduran-2-sulfonylamino)-butyricacid

(L)-3-Methyl-2-(7-phenyl-dibenzofuran-2-sulfonylamino)-butyric acid,tert.-butyl ester (0.94 g, mmoles) was dissolved in concentratedtrifluoroacetic acid and stirred for 2 hours. Concentrated in vacuo andtriturated the residue with diethyl ether to give the title compound asan off-white solid; melting point=254-255° C.

Inhibition Studies

Experiments were carried out which demonstrate the efficacy of compoundsof Formula I and II as potent inhibitors of stromelysin-1 and gelatinaseA. Experiments were carried out with the catalytic domains, i.e., Table1 shows the activity of the Examples with respect to both stromelysin-1and gelatinase A, GCD (recombinant gelatinase A catalytic domain); SCD(stromelysin-1 catalytic domain). IC₅₀ values were determined using athiopeptolide substrate, Ac-Pro-Leu-Gly-thioester-Leu-Leu-Gly-OEt (YeQ.-Z., Johnson L. L., Hupe D. J., and Baragi V., “Purification andCharacterization of the Human Stromelysin Catalytic Domain Expressed inEscherichia coli,” Biochemistry, 1992;31:11231-11235). MMP01, MMP07,MMP09, and MMP13 activity was assayed in a method similar to MMP02 andMMP03 (SCD and GCD). MMP01 and MMP09 can be obtained from WashingtonUniversity School of Medicine, St. Louis, Mo. MMP07 can be obtained inaccordance with the known procedure set forth by Ye Q-Z, Johnson L. L.,and Baragi V., “Gene Syntheses and Expression in E. coli for PUMP, aHuman Matrix Metalloproteinase” Biochem. and Biophys. Res. Comm.,1992;186:143-149. MMP13 can be obtained in accordance with the knownprocedure set forth by Freije J. M. P., et al., “Molecular Cloning andExpression of Collegenase-3, a Novel Human Matrix MetalloproteinaseProduced by Breast Carcinomas” J. Bio. Chem., 1994;269:16766-16773.

Thiopeptolide Assay

Hydrolysis of the thiopeptolide substrateAc-Pro-Leu-Gly-thioester-Leu-Leu-Gly-OEt (Bachem) is used as the primaryscreen to determine IC₅₀ values for MMP inhibitors. A 100 μL reactioncontains 1 mM 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), 100 μMsubstrate, 0.1% Brij, enzyme, and inhibitor in the appropriate reactionbuffer. Activated full-length enzymes are assayed at 5 nM, StromelysinCatalytic Domain (SCD) at 10 nM, and Gelatinase A Catalytic Domain(GaCD) at 1 nM. Inhibitors are screened from 100 μM to 1 nM. Full-lengthenzymes are assayed in 50 mM HEPES, 10 mM CaCl₂, pH 7.0; SCD in 50 mMMES, 10 mM CaCl₂, pH 6.0; and GaCD in 50 mM MOPS, 10 mM CaCl₂, 10 μMZnCl₂, pH 7.0. The change in absorbance at 405 nm is monitored on aThermoMax microplate reader at room temperature continuously for 20minutes.

HEPES is 4-(2-hydroxylethyl)-piperazine-1-ethane sulfonic acid;

MES is 2-morpholinoethane sulfonic acid menohydrate;

Ac is acetyl;

Pro is proline;

Leu is leucine;

Gly is glycine;

Et is ethyl; and

MOPS is 3-morpholinopropane sulfonic acid.

Soluble Proteoglycan Assay (Stromelysin Natural Substrate Assay) SCD(PG)

Solubilized proteoglycan substrate is prepared from bovine cartilagepowder (Sigma) using the method described by Nagase and Woessner inAnal. Biochem., 1980;107:385-392. A 100 μL reaction contains 10 μg/mLproteoglycan, enzyme, and inhibitor in 50 mM MES, 10 mM CaCl₂, pH 6.0.Activated full-length stromelysin or stromelysin catalytic domain (SCD)is assayed at 100 nM. Inhibitors are screened from 100 μM to 1 nM. Thereaction is incubated at 37° C. for 3 hours then stopped with theaddition of 1,10-phenanthroline at a final concentration of 1 mM.Reaction products are separated from undigested substrate usingultrafree-MC polysulfone microcons with a 300,000 molecular weightcut-off membrane (Millipore) and quantified using a modified1,9-dimethylene blue (DMB) assay described by Farndale, Sayers, andBarrett in Connective Tissue Research, 1982;9:247-248. Absorbance ismeasured at 518 nm using 32 μg/mL DMB in a 1 mL reaction. The standardcurve is constructed from 0 to 100 μg shark cartilage chondroitinsulfate C (Sigma).

Gelatin Assay (Gelatinase Natural Substrate Assay) (Gel)

Rat tail Type I collagen (Sigma) is denatured by heating at 95° C. for20 minutes to prepare the gelatin substrate. A 50 μL reaction contains1.12 mg/mL substrate, enzyme, inhibitor, and 80 μg/mL soy bean trypsininhibitor as an inert internal standard in 50 mM MOPS, 10 mM CaCl₂, 10μM ZnCl₂, pH 7.0. Activated full-length gelatinase A is assayed at 1 nMand gelatinase A catalytic domain (GaCD) at 10 nM. Inhibitors arescreened from 100 μM to 1 nM. The reactions are incubated at 37° C. for30 minutes then stopped with 50 μL at 2× Tricine gel loading buffer(Novex). Reaction products are separated from undigested substrate byelectrophoresis on Tricine-SDS 10-20% polyacrylamide gradient gels(Novex). Protein bands are stained with Coomassie Brilliant Blue R andquantified using a Bio Image densitometer (Millipore). IC₅₀ values arecalculated from the disappearance of substrate using the sum of the topthree bands of each reaction after normalization with the internalstandard.

MMP Inhibitor Bioassay

Animals are dosed by gavage with either vehicle or compound at 2, 10, or50 mg/kg. Blood samples are collected from 3 to 4 animals from eachdosing group at 1, 2, 4, 6, and 24 hour postdose, centrifuged, and theplasma immediately frozen at −20° C. Plasma protein is precipitated withan equal volume of acetonitrile and separated by centrifugation at roomtemperature. The supernate is evaporated to dryness and reconstituted tothe original plasma volume with 50 mM Tris, pH 7.6. Ten-fold serialdilutions of the reconstituted plasma samples are prepared in 50 mMTris, pH 7.6 for dose response assays using the appropriatethiopeptolide assay. The concentration of plasma which yields 50%inhibition of enzyme is determined and used to calculate the inhibitorplasma level from the known IC₅₀ value. To demonstrate that the compoundcan be quantitatively extracted from plasma as active inhibitor,controls for each inhibitor include normal rat plasma, normal rat plasmaspiked with compound, and buffer dilutions of compound. All controlsamples are subjected to acetonitrile precipitation and analyzed withthe thiopeptolide assay.

TABLE 1 Example (GCD) (SCD) Number MMP01 MMP02 MMP03 MMP07 MMP09 MMP13 166 0.32 1.18 — 100 — 2 100 2.3 1.5 — 100 — 3 100 0.9 0.72 — 100 — 4 —1.7 5.4 — — — 5 19 0.084 0.23 — 100 — 6 100 0.73 4.8 — 100 — 7 — 1.2 1.0— — — 8 — 9.4 14.4 — — — 9 — 4.5 0.69 — — — 10 — 35 100 — — — 11 1.80.0045 0.015 5.0 — 0.047 12 32.3 0.049 0.185 10.8 100 0.34 13 — — — — —— 14 100 0.61 0.69 27 — 2.6 15 100 100 100 100 — 100 16 — 0.47 0.75 — —— 17 100 0.36 0.062 6 — 0.69

TABLE 2 Example 5 SCD (IC₅₀) 0.233 μM SCD (PG) (IC₅₀) 8.9 μM GCD (IC₅₀)0.084 μM Gel (IC₅₀) 0.58 μM Bioassay (50 mg/kg) Peak 82 μM 24 Hours 0.18μM

1. A method of treating multiple sclerosis, the method comprisingadministering to a patient having multiple sclerosis a therapeuticallyeffective amount of a compound of Formula I

wherein M is a natural (L) alpha amino acid derivative having thestructure

X is O, S, S(O),S(O)₂, CH₂, CO, or NR^(Q); R^(Q) is hydrogen, C₁-C₆alkyl, or —C₁-C₆ alkyl-phenyl; R is a side chain of a natural alphaamino acid; R¹ is C₁-C₅ alkoxy, hydroxy, or —NHOR⁵; R² and R⁴ areindependently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂, halogen, —OR⁵, —CN,—CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶, —(CH₂)_(n)NR⁵R⁶, —CF₃, or—NHCOR⁵; each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl; and nis 0 to 2, and the pharmaceutically acceptable salts, esters, and amidesthereof, wherein the esters thereof are selected from C₁-C₆ alkylesters, C₅-C₇ cycloalkyl esters, and arylalkyl esters and the amidesthereof are derived from ammonia, primary C₁-C₆ alkyl amines, secondaryC₁-C₆ dialkyl, and 5- and 6-membered hererocyclic amines containing onenitrogen atom; and wherein the group S(═O)₂M is optionally bonded to the1-, 2-, or 3-position of Formula I.
 2. A method of treating arthritis,the method comprising administering to a patient having arthritis atherapeutically effective amount of a compound of Formula I

wherein M is a natural (L) alpha amino acid derivative having thestructure

X is O, S, S(O), S(O)₂, CH₂, CO, or NR^(Q); R^(Q) is hydrogen, C₁-C₆alkyl, or —C₁-C₆ alkyl-phenyl; R is a side chain of a natural alphaamino acid; R¹ is C₁-C₅ alkoxy, hydroxy, or —NHOR⁵; R² and R⁴ areindependently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂, halogen, —OR⁵) , —CN,—CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶, —(CH₂)_(n)NR⁵R⁶, —CF₃, or—NHCOR⁵; each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl; and nis 0 to 2, and the pharmaceutically acceptable salts, esters, and amidesthereof, wherein the esters thereof are selected from C₁-C₆ alkylesters, C₅-C₇ cycloalkyl esters, and arylalkyl esters and the amidesthereof are derived from ammonia, primary C₁-C₆ alkyl amines, secondaryC₁-C₆ dialkyl, and 5- and 6-membered heterocyclic amines containing onenitrogen atom; and wherein the group S(═O)₂M is optionally bonded to the1-, 2-, or 3-position of Formula I.
 3. A compound of Formula I

wherein M is a natural (L) alpha amino acid derivative having thestructure

X is S, S(O), S(O)₂, CO, or NR^(Q); R^(b) is a side chain of a naturalalpha amino acid; R^(a) is C₁-C₅ alkoxy, hydroxy, or —NHOR⁵; R² and R⁴are independently hydrogen, —C₁-C₅ alkyl, phenyl —NO₂, halogen, —OR⁵,—CN, —CO₂R⁵, —SO₃R⁵, —CHO, —COR⁵, —CONR⁵R⁶, —(CH₂)_(n)NR⁵R⁶,—CF₃, or—NHCOR⁵; each R⁵ and R⁶ are independently hydrogen or C₁-C₅ alkyl; and nis 0 to 2, and the pharmaceutically acceptable salts, esters, and amidesthereof, wherein the esters thereof are selected from C₁-C₆ alkylesters, C₅-C₇ cycloalkyl esters, and arylalkyl esters and the amidesthereof are derived from ammonia, primary C₁-C₆ alkyl amines, secondaryC₁-C₆ dialkyl, and 5- and 6-membered heterocyclic amines containing onenitrogen atom; and wherein the group S(═O)₂M is optionally bonded to the1-, 2-, or 3-position of Formula I.